• Title/Summary/Keyword: Small protein

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Impact of Sodium Copper Chlorophyllin on Mercury Absorption Using an in Vitro Digestion with Human Intestinal Cell Model

  • Hwang, Han-Joon;Shim, Soon-Mi
    • Food Science and Biotechnology
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    • v.17 no.3
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    • pp.564-568
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    • 2008
  • The effects of sodium copper chlorophyllin (SCC) on bioaccessibility and uptake of mercury from fish were investigated using an in vitro digestion coupled with a Caco-2 cell. Fish along with SCC was subjected to a simulated in vitro digestion, which simulates both the gastric and small intestinal phase in vivo. Mercury bioaccessibility, the amount of mercury released from fish to aqueous phase following a digestion, was measured. Various amounts of SCC (0.1-25 mg) significantly reduced mercury bioaccessibility in a dose dependent manner by 49-89% compared to the negative control (fish without SCC) (p<0.05). Mercury bioaccessibility in varying molar ratios of mercury to positive control, 2,3-dimercapto-1-propane sulfonate (DMPS) was between 24 and 52%. Mercury uptake by Caco-2 cells from test media containing aqueous phase following in vitro digestion was measured after 6 hr incubation at $37^{\circ}C$. Cellular mercury uptake with increasing amount of SCC ranged from 0.352 to $0.052\;{\mu}g$ mercury/mg protein, while those in DMPS treatment were between 0.14 and $0.27\;{\mu}g$ mercury/mg protein. Our study suggests that SCC can reduce mercury absorption following fish consumption and may be efficient as a synthetic chelating agent for long term chronic mercury exposure in fish eating populations.

Construction of an Oscillator Gene Circuit by Negative and Positive Feedbacks

  • Shen, Shihui;Ma, Yushu;Ren, Yuhong;Wei, Dongzhi
    • Journal of Microbiology and Biotechnology
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    • v.26 no.1
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    • pp.139-144
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    • 2016
  • Synthetic oscillators are gene circuits in which the protein expression will change over time. The delay of transcription, translation, and protein folding is used to form this kind of behavior. Here, we tried to design a synthetic oscillator by a negative feedback combined with a positive feedback. With the mutant promoter PLacC repressed by LacIq and PLux activated by AHL-bound LuxR, two gene circuits, Os-LAA and Os-ASV, were constructed and introduced into LacI-deleted E. coli DH5α cells. When glucose was used as the carbon source, a low level of fluorescence was detected in the culture, and the bacteria with Os-ASV showed no oscillation, whereas a small portion of those carrying Os-LAA demonstrated oscillation behavior with a period of about 68.3 ± 20 min. When glycerol was used as the carbon source, bacteria with Os-ASV demonstrated high fluorescence value and oscillation behavior with the period of about 121 ± 21 min.

A Receptor Tyrosine Kinase Inhibitor, Dovitinib (TKI-258), Enhances BMP-2-Induced Osteoblast Differentiation In Vitro

  • Lee, Yura;Bae, Kyoung Jun;Chon, Hae Jung;Kim, Seong Hwan;Kim, Soon Ae;Kim, Jiyeon
    • Molecules and Cells
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    • v.39 no.5
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    • pp.389-394
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    • 2016
  • Dovitinib (TKI258) is a small molecule multi-kinase inhibitor currently in clinical phase I/II/III development for the treatment of various types of cancers. This drug has a safe and effective pharmacokinetic/pharmacodynamic profile. Although dovitinib can bind several kinases at nanomolar concentrations, there are no reports relating to osteoporosis or osteoblast differentiation. Herein, we investigated the effect of dovitinib on human recombinant bone morphogenetic protein (BMP)-2-induced osteoblast differentiation in a cell culture model. Dovitinib enhanced the BMP-2-induced alkaline phosphatase (ALP) induction, which is a representative marker of osteoblast differentiation. Dovitinib also stimulated the translocation of phosphorylated Smad1/5/8 into the nucleus and phosphorylation of mitogen-activated protein kinases, including ERK1/2 and p38. In addition, the mRNA expression of BMP-4, BMP-7, ALP, and OCN increased with dovitinib treatment. Our results suggest that dovitinib has a potent stimulating effect on BMP-2-induced osteoblast differentiation and this existing drug has potential for repositioning in the treatment of bone-related disorders.

Different expression of human GFAP promoter-derived GFP in different subsets of astrocytes in the mouse brain

  • Moon, Young-Hye;Kim, Hyun-Jung;Kim, Joo-Yeon;Kim, Hyun;Kim, Woon-Ryoung;Sun, Woong
    • Animal cells and systems
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    • v.15 no.4
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    • pp.268-273
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    • 2011
  • Transgenic mice expressing green fluorescent protein (GFP) under the control of human glial fibrillary acidic protein promoter (hGFAP) have been utilized for in vivo labeling of astrocytes. Although it has been considered that virtually all astrocytes express GFP in this transgenic mouse, we found that different subsets of GFAP-expressing astrocytes express considerably different levels of GFP in the adult brain. Astrocytes in the spinal cord, the molecular layer of thecerebellum, meninges, white matter, corpus callosum and blood vessels exhibited strong GFP, whereas subsets of astrocytes associated with granule cells in the cerebellum and dentate gyrus did not or only marginally exhibited GFP. We also found that a small subset of GFP-expressing cells in the periglomeruli of the olfactory bulb did not express GFAP immunoreactivity. Collectively, these results suggest that human GFAP promoter-derived GFP expression does not faithfully recapitulate the endogenous GFAP expression in mice, suggesting that upstream regulatory mechanisms controlling GFAP transcription are different in different populations of astrocytes, and may reflect the functional diversity of astrocytes.

Influence of Caecectomy on the Bioavailability of Minerals from Vegetable Protein Supplements in Adult Roosters

  • Vasan, P.;Dutta, Narayan;Mandal, A.B.;Sharma, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.8
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    • pp.1178-1182
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    • 2008
  • The present study was designed to assess the influence of caeca on the availability of calcium, phosphorus, magnesium, manganese and copper from soybean, sunflower, rapeseed, sesame, fish and meat cum bone meal in adult roosters. The excretion of endogenous origin minerals viz., copper, magnesium, manganese and calcium was significantly (p<0.001) higher in caecectomized than in normal roosters. The difference in the endogenous excretion was 50; 60.45; 40.35 and 29.63 per cent for copper, magnesium, manganese and calcium, respectively, in caecectomized roosters. The caeca played a pivotal role in the reabsorption of endogenous origin calcium, magnesium, manganese and copper. The mechanism of phosphorus absorption by the caecal epithelium was negligible. The caecectomized roosters underestimated the bioavailability of copper in sunflower meal and manganese in almost all the test feedstuffs. The present investigation revealed that the caeca played a critical role in the absorption of minerals from vegetable protein feedstuffs which escape digestion and absorption in the small and large intestinal segments.

Anesthetic-Induced Acid-Base Equilibrium and Its Electric Dichroism on the Bacteriorhodopsin (Bacteriorhodopsin에 관한 마취제 유도 산-염기 평형 및 Electric Dichroism)

  • Lee, Ki Hwan
    • Journal of the Korean Chemical Society
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    • v.42 no.2
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    • pp.143-149
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    • 1998
  • When anesthetics were added to a bacteriorhodopsin (bR) in the purple membranes, the 570 nm absorption band shifts to 480 nm. This anesthetic-induced spectroscopic change is reversible. The apparent pKa (6.3) of this equilibrium depends on the nature of the anesthetics in which bR is dispersed. The electric orientation measurements showed that the native bR is easily achieved by relatively small electric field which is oriented at $60^\circ$, while anesthetic-treated bR is not the case. These results demonstrate that the subtle changes in the chromophore and the protein structure surrounding the chromophore by anesthetics influence the spatial orientation of the charged residues in the protein matrix surrounding chromophore.

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Purification of Deoxycytidine Kinase from Various Human Leukemic Cells by End-product Analog Affinity Chromatography

  • Kim, Min-Young
    • BMB Reports
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    • v.28 no.4
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    • pp.281-289
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    • 1995
  • Homogeneous human deoxycytidine kinase was purified in one step from a variety of spontaneous human leukemic cells (T-ALL, B-ALL, B-CLL, AML, CML), and from cultured T-lymphoblast cells (MOLT-4) using the newly developed affinity medium, $dCp_4$-Sepharose. Starting with an ammonium sulfate fraction, purification was achieved in one step with the kinase being eluted from a column by the end product inhibitor, dCTP. The purified deoxycytidine kinase from T-ALL cells phosphorylated deoxyadenosine and deoxyguanosine, as well as deoxycytidine. The enzyme purified from T-ALL and B-CLL cells yielded one major band with a molecular weight of 52 kDa determined by SDS-polyacrylamide gel electrophoresis. AML and CML cells yielded one 52 kDa band and an extra band of 30 kDa molecular weight. On the other hand, B-ALL and MOLT-4 cells showed a low molecular weight band of 30 kDa only. However, the electrophoretic mobilities of enzymatic activity in 12% non-denaturing gels were identical for the dCyd kinase from all different kinds of leukemic cell lines, except that the B-ALL, B-CLL, and MOLT-4 cell preparations had an extra minor peak, all at the same position. dAdo and dCyd phosphorylating activities comigrated indicating that these activities are all associated with the same protein. Two new methods, a disk implantation method and a nitrocellulose powder method were used with a small amount of enzyme protein to raise polyclonal antibodies against dCyd kinase purified from T-ALL cells.

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Transcriptome Analysis to Characterize the Immune Response of NecroX-7 in Mouse CD4+ T Cells

  • Kim, Eun-Jung
    • Biomedical Science Letters
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    • v.21 no.2
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    • pp.60-68
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    • 2015
  • NecroX-7 is a novel small compound of the NecroX series based on the indole moiety, which has potent cytoprotective and antioxidant properties. We previously detected potential immune regulatory effects of NecroX-7 in immune related diseases like Graft-versus-Host Disease. However, the function and the underlying mechanisms of immunological effects of NecroX-7 in the immune system have not been well established. In this study, we investigated the immune response characterization of differentially expressed genes of NecroX-7 administration in $CD4^+$ T cells by microarray analysis. $CD4^+$ T cells stimulated with NecroX-7 ($40{\mu}M$) or vehicle for 72 hours resulted in the identification of 337 differentially expressed genes (1.5 fold, P<0.05) by expression profiling analysis. Twenty eight of the explored NecroX-7-regulated genes were related to immune system processes. These genes were validated by quantitative real-time PCR. The most significant genes were glutathione reductase, eukaryotic translation elongation factor 1, lymphotoxin-alpha, heat shock protein 9 and chloride intracellular channel protein 4. These findings demonstrate the strongly immune response of NecroX-7 in $CD4^+$ T cells, suggesting that cytoprotection and immune regulation may underlie the critical aspects of NecroX-7 exposure.

Adjuvant Therapy Efficacy of Herbal Medicine Zeo Lyung Tang (Zhu Ling Decoction) for Primary Glomerulonephritis: Systematic Review and Meta-Analysis (일차성 사구체신염에 대한 저령탕 병용요법의 효능 : 체계적 문헌고찰 및 메타 분석)

  • Kim, Bomin;Jo, Hee-Geun
    • The Journal of Internal Korean Medicine
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    • v.41 no.4
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    • pp.644-657
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    • 2020
  • Objectives: The aim of this study was to systematically evaluate the clinical therapeutic effects and safety of the Zeo lyung tang (ZLT) on primary glomerulonephritis (PGN). Methods: The MEDLINE, EMBASE, PubMed, CENTRAL, CNKI, RISS, NDSL, KISS, and OASIS databases were searched for randomized controlled trials (RCTs) testing the effects of ZLT on PGN. The Cochrane collaboration bias risk assessment scale was used to evaluate the methodological quality of the included studies. RevMan 5.3 software was used for data analysis. Results: Ten RCTs involving 781 patients were included in the review. Compared with conventional Western medicine (WM) therapy alone, a combination treatment of ZLT and WM improved the total effective rate (RR=1.24; 95%CI [1.16, 1.33]; p<0.00001), reduce the blood urea nitrogen (BUN; MD=-1.05; 95%CI [-1.32, -0.78]; p<0.00001) and the 24-hour urinary protein (MD=-0.38; 95%CI [-0.46, -0.29]; p<0.00001). Conclusions: The combination of ZLT with WM has therapeutic effects on PGN, and it has advantages over WM treatment alone in reducing BUN and 24-hour urinary protein. However, due to the low quality of the included studies and the small sample sizes, additional research is needed in this area.

Stress Responses through Heat Shock Transcription Factor in S. cerevisiae

  • Hahn, Ji-Sook;Hu, Zhanzhi;Thiele, Dennis J.;Lyer, Vishwanath R.
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2005.05a
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    • pp.105-109
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    • 2005
  • Heat Shock Transcription Factor (HSF), and the promoter heat Shock Element (HSE), are among the most highly conserved transcriptional regulatory elements in nature. HSF mediates the transcriptional response of eukaryotic cells to heat, infection and inflammation, pharmacological agents, and other stresses. While HSF is essential for cell viability in yeast, oogenesis and early development in Drosophila, extended life-span in C. elegans, and extra-embryonic development and stress resistance in mammals, little is known about its full range of biological target genes. We used whole genome analyses to identify virtually all of the direct transcriptional targets of yeast HSF, representing nearly three percent of the genomic loci. The majority of the identified loci are heat-inducibly bound by yeast HSF, and the target genes encode proteins that have a broad range of biological functions including protein folding and degradation, energy generation, protein secretion, maintenance of cell integrity, small molecule transport, cell signaling, and transcription. Approximately 30% of the HSF direct target genes are also induced by the diauxic shift, in which glucose levels begin to be depleted. We demonstrate that phosphorylation of HSF by Snf1 kinase is responsible for expression of a subset of HSF targets upon glucose starvation.

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